1. Influence of Protein Scaffold on Side-Chain Transfer Free Energies
Dagen C. Marx, Karen G. Fleming 
Biophysical Journal 
Volume 113, Issue 3, Pages (August 2017)
DOI: /j.bpj
Copyright © 2017 Biophysical Society Terms and Conditions

2. Figure 1
Comparison of structural characteristics of PagP and OmpLA. PagP (PDB: 1MM4) (left) is colored in cyan and has eight transmembrane β-strands and a periplasmic helix. OmpLA (PDB: 1QD5) (right), colored green, has 12 transmembrane strands and is not tilted with respect to the bilayer (shown as black lines). Sites at which transfer free energies were measured are shown as black spheres (PagP V111 and OmpLA A210), highlighting the different depths of the two positions. Shown below the structures are the nearest-neighbor residues of the sites mutated in the two studies (shown in black spheres). This figure was created using PyMOL (33). To see this figure in color, go online.
Biophysical Journal  , DOI: ( /j.bpj )
Copyright © 2017 Biophysical Society Terms and Conditions

3. Figure 2
All PagP variants show path independence in chemical denaturation titrations. Unfolding titrations are shown by open symbols and overlaid upon folding titrations, which are shown by solid symbols. The variant is noted in each box, and the coloring scheme is identical to that in Fig. 3 and Fig. S1. To see this figure in color, go online.
Biophysical Journal  , DOI: ( /j.bpj )
Copyright © 2017 Biophysical Society Terms and Conditions

4. Figure 3
High-resolution chemical denaturation titrations of PagP variants. Representative chemical denaturation titration data for all 20 PagP variants at position V111 with ionizable residues colored blue, polar residues red, nonpolar residues black, and cysteine and glycine in gold. Normalized intrinsic tryptophan fluorescence intensity is plotted as a function of the concentration of guanidine HCl. Data were fit to a two-state folding model with the m-value held constant (m = 4.97). To see this figure in color, go online.
Biophysical Journal  , DOI: ( /j.bpj )
Copyright © 2017 Biophysical Society Terms and Conditions

5. Figure 4
Determination of the nonpolar solvation parameter at PagP site 111. The ΔΔGw,lo values of nonpolar residues were plotted as a function of their nonpolar accessible surface area in a G-X-Gly tripeptide, where X is any amino acid. The correlation is well described (R2 = 0.91), with intercept equal to and slope equal to 25 cal mol−1 A−2, which reflects the value of the σNP from bilayer to water.
Biophysical Journal  , DOI: ( /j.bpj )
Copyright © 2017 Biophysical Society Terms and Conditions

6. Figure 5
Reference-free ΔΔGsco values for all side chains. Side-chain transfer free energies for all 20 amino acids at site 111 after correcting for the transfer free energy of alanine. Error bars are standard deviations (n = 3 for all variants). To see this figure in color, go online.
Biophysical Journal  , DOI: ( /j.bpj )
Copyright © 2017 Biophysical Society Terms and Conditions

7. Figure 6
V111P induces local unfolding in PagP. (A) To assess the effect of the V111P variant on PagP structure we measured the distance between the backbone hydrogen-bonding atoms between residues 111 and 85. The top panel shows the distance between the carbonyl oxygen on residue 111 and the backbone nitrogen of residue 85, with the two WT trajectories colored gray and V111P trajectories colored red. The bottom panel shows the distance between the residue 111 backbone nitrogen and the residue 85 carbonyl oxygen, with WT PagP trajectories colored gray and V111P trajectories colored blue. The distances in V111P are too large for any contacts between the residues indicating a local unfolding event. (B) Snapshot from V111P trajectory showing the local unfolding of the β-sheet. The coloring scheme from (A) is used to show the two different distances being measured. (C) Snapshot from the WT trajectory showing close contacts between residues 111 and 85. (B) and (C) were created using PyMoL, and DLPC phosphates are shown as orange spheres in both images (33). To see this figure in color, go online.
Biophysical Journal  , DOI: ( /j.bpj )
Copyright © 2017 Biophysical Society Terms and Conditions

8. Figure 7
PagP and OmpLA hydrophobicity scales are well correlated. The plot shows the comparison of the two whole-protein water-to-bilayer hydrophobicity scales with PagP on the y axis and OmpLA on the x axis. Residues are colored with the same coloring scheme as in Fig. 2. The 95% confidence intervals of the fit are shown in light blue. Proline was excluded from the linear fit and subsequent determination of confidence intervals. To see this figure in color, go online.
Biophysical Journal  , DOI: ( /j.bpj )
Copyright © 2017 Biophysical Society Terms and Conditions